Recently, Chu Jun, the research team of the Biomedical Optics and Molecular Imaging Research Center of the Institute of Biomedical and Health Engineering (hereinafter referred to as the Institute of Biomedical Engineering) of the Shenzhen Advanced Technology Research Institute of the Chinese Academy of Sciences (hereinafter referred to as the Shenzhen Advanced Technology Research Institute), developed a high-performance gene coded cyclic adenosine monophosphate (cAMP) green fluorescent probe (G-Flamp1) with 12 times fluorescence changes in living cells. This study combines microscopic imaging and optical fiber recording technologies to detect the spatiotemporal dynamics of cyclic adenosine monophosphate signal of specific neurons in the specific behavior process of model organisms such as drosophila and mice in a real-time and highly sensitive manner, and explores the internal relationship between cyclic adenosine monophosphate dynamics and animal behavior. Relevant research papers were published recently in Nature Communication. Cell is the basic unit of structure and function of most living things, including human beings. Cells will constantly receive signals from the surrounding environment and correspondingly change the quantity, distribution and active state of proteins, small organic molecules, ions, DNA and RNA in cells, thus changing their own morphology and biological functions. The abnormality of this process is related to the occurrence and development of the disease. Therefore, scientists often clarify the pathogenesis of related diseases by detecting the spatiotemporal changes of the above key molecules. In this study, the researchers selected adenosine cyclic phosphate, an important second messenger molecule in cells, as the research target. Cyclic adenosine monophosphate can transfer the information of various G protein coupled receptors (GPCR) on the cell surface, and plays an important role in learning and memory, drug addiction, motor control, immunity, metabolism and other processes. "High spatial and temporal resolution fluorescence imaging of changes in the concentration of cyclic adenosine monophosphate molecules at the living cell and living level is an important basis for analyzing the cyclic adenosine monophosphate signal pathway and its biological functions. Therefore, the development of highly sensitive cyclic adenosine monophosphate fluorescent probes has become the key to the study of complex biological processes." Chu Jun said. Compared with non gene coding probes, gene coding probes, like normal proteins, can locate specific cells or specific cell substructures of organisms. They have the advantages of low toxicity, low background, heritability, etc., and have unparalleled advantages in basic research of life science. However, the existing adenosine cyclophosphate fluorescent probes encoded by more than 50 genes are either low sensitivity (the maximum fluorescence change is only 1.5 times) or low fluorescence brightness, which makes it difficult to monitor the weak endogenous adenosine cyclophosphate changes in vivo, greatly limiting the research on the molecular regulatory mechanism and function of adenosine cyclophosphate under physiological and pathological conditions. In order to develop a highly sensitive probe for in vivo detection, cyclically rearranged green fluorescent protein (cpGFP) was inserted into the cyclic adenosine phosphate binding domain (mlCNBD) of bacterial MlotiK1 channel protein. After selection of insertion sites, optimization of linker peptides, optimization of fluorescent proteins and sensing modules, researchers have obtained high-performance gene coded adenosine cyclic phosphate green fluorescent probe (G-Flamp1) with high brightness, high sensitivity, appropriate affinity and fast response speed. The fluorescence change of the probe in living cells can reach 12 times, which is one of the few fluorescent probes with fluorescence change of more than 10 times. Later
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